SCIENTIFIC OPINION. EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS) 2, 3. European Food Safety Authority (EFSA), Parma, Italy

SCIENTIFIC OPINION Scientific Opinion on the safety of sucrose esters of fatty acids prepared from vinyl esters of fatty acids and on the extension of use of sucrose esters of fatty acids in flavourings 1 ABSTRACT EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS) 2, 3 European Food Safety Authority (EFSA), Parma, Italy The Panel on Food Additives and Nutrient Sources added to Food provides a scientific opinion on the safety of sucrose esters of fatty acids produced by a new manufacturing method from sucrose and vinyl esters of fatty acids, and an evaluation of the extension of the use of this additive to flavoured fruit beverages. In 2004 EFSA established a group ADI of 40 mg/kg bw/day for sucrose esters of fatty acids, however this evaluation did not consider sucrose esters of lauric acid. The Panel considers that although the toxicological data on the sucrose ester of lauric acid are limited, they do not give rise to concerns since lauric acid is a natural constituent of a number of foods. The new manufacturing method results in residues of vinyl esters of fatty acids of <10-111 mg/kg and acetaldehyde of 20-48 mg/kg in the sucrose esters of fatty acids. p-methoxyphenol was undetectable. The vinyl portion of the vinyl ester instantly tautomerises to acetaldehyde in the gastrointestinal tract. The Panel concludes that the additional average exposure of 1.4 µg acetaldehyde/kg bw/day resulting from the use of sucrose esters of fatty acids as a food additive would be negligible compared to the exposure from food and endogenous formation and not of safety concern. The Panel also concludes that since p- methoxyphenol was undetectable, this potential residue is not of safety concern. The Panel therefore concludes that sucrose esters of fatty acids produced by the new manufacturing method do not present any safety concern provided the overall exposure is within the ADI of 40 mg/kg bw/day. The Panel notes that the current intake of sucrose esters of fatty acids is high and for some individuals above the ADI but that an additional intake from fruit beverages only seems to contribute to a few percent of the ADI. KEY WORDS Sucrose monoesters of fatty acids, manufacturing method, vinyl esters of fatty acids, acetaldehyde, E473, food additive, emulsifier, stabiliser 1 On request from the European Commission, Question No EFSA-Q-2009-00451, adopted on 10 February 2010. 2 Panel members: F. Aguilar, U.R. Charrondiere, B. Dusemund, P. Galtier, J. Gilbert, D.M. Gott, S. Grilli, R. Guertler, J. Koenig, C. Lambré, J-C. Larsen, J-C. Leblanc, A. Mortensen, D. Parent-Massin, I. Pratt, I.M.C.M. Rietjens, I. Stankovic, P. Tobback, T. Verguieva, R.A. Woutersen. Correspondence: ans@efsa.europa.eu 3 Acknowledgement: The Panel wishes to thank the members of the Working Group A on Food Additives and Nutrient Sources of the ANS Panel for the preparation of this opinion: F. Aguilar, N. Bemrah, P. Galtier, J. Gilbert, S. Grilli, R. Guertler, NG. Ilback, C. Lambré, J.C. Larsen, J-C. Leblanc, A. Mortensen, I. Pratt, Ch. Tlustos, I. Stankovic. Suggested citation: EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS); Scientific Opinion on the safety of sucrose esters of fatty acids prepared from vinyl esters of fatty acids and on the extension of use of sucrose esters of fatty acids in flavourings on request from the European Commission. EFSA Journal 2010;(3):1512. [36 pp.]. doi:10.2903/j.efsa.2010.1512. Available online: www.efsa.europa.eu European Food Safety Authority, 2010 1

SUMMARY Following a request from the European Commission to the European Food Safety Authority (EFSA), the Scientific Panel on Food Additives and Nutrient Sources added to Food (ANS) was asked to deliver a scientific opinion on the safety in use as food additive of sucrose esters of fatty acids produced from an alternative route by reacting sucrose and vinyl esters of fatty acids. In addition the applicant has requested an extension of the use of sucrose esters of fatty acids to allow the additives to be used in flavourings in order to improve solubility of the flavouring in water based beverages. Sucrose esters of fatty acids (E 473) are authorised for use in a number of foods via Directive 95/2/EC of the European Parliament and Council on food additives other than colours and sweeteners. Sucrose esters of fatty acids were evaluated by the EC Scientific Committee for Food (SCF) in 1992. At that time the SCF established a group ADI of 0-20 mg/kg bw/day (expressed as sucrose monostearate) for sucrose esters of fatty acids and sucroglycerides derived from palm oil, lard and tallow fatty acids. In 2004, in light of new studies which had been provided, EFSA re-examined the safety of these food additives and established a group ADI of 40 mg/kg bw/day for sucrose esters of fatty acids (E 473) and sucroglycerides (E 474). However, sucrose ester of lauric acid were not considered in these evaluations. Sucrose esters of fatty acids (and sucroglycerides) were evaluated by JECFA in 1992 and 1995. In the later evaluation JECFA allocated a temporary group ADI of 0-20 mg/kg bw/day (WHO, 1995). JECFA made the ADI temporary and requested the results of a well designed and conducted tolerance study for review in 1997 (WHO, 1995). This study was submitted and evaluated by JECFA in 1997 and a full group ADI of 0-30 mg/kg bw was established (WHO, 1998). In 2009, JECFA established a group ADI of 0-30 mg/kg bw/day for sucrose esters of fatty acids, sucroglycerides and sucrose oligoesters type I and type II (JECFA, 2009). The present opinion deals with the safety of sucrose esters of fatty acids (monoesters of lauric acid, myristic acid, palmitic acid, and stearic acid) produced by an alternative route by reacting sucrose and vinyl esters of fatty acids. The new manufacturing process results in residue levels of vinyl esters of fatty acid from <10-111 mg/kg, acetaldehyde (formed from the vinyl portion of the vinyl fatty acid esters) levels from 20-48 mg/kg, and not detectable (<0.1 mg/kg) levels of p-methoxyphenol (stabiliser) in the sucrose esters of fatty acids manufactured by this process. In addition, the opinion has considered whether a requested extension of the use of sucrose esters of fatty acids in flavourings in water based beverages will increase the total intake of sucrose esters of fatty acids. The Panel concludes that these monoesters are extensively hydrolysed in the gastrointestinal tract into the constituent fatty acids and sucrose prior to absorption. The Panel considers that there is no reason to believe that the sucrose monoesters of fatty acids per se produced by the new manufacturing process should in any way have biological or toxicological effects different from those of sucrose monoesters of fatty acids produced by the currently-used manufacturing methods. The Panel notes that lauric acid is a natural dietary constituent found at relatively high concentrations in a number of foods, but that no specific toxicological data are available on the sucrose ester of lauric acid. The Panel considers that although the available data on the toxicological profile of lauric acid are limited, they do not give rise to specific concerns. The maximum residual level of vinyl esters of fatty acids in sucrose esters of fatty acids, as reported by the petitioner, is around 111 mg/kg. The daily average and high level (95 th percentile) exposure of Irish adults to vinyl fatty acid esters from all foodstuffs, in which sucrose esters of fatty acids are permitted, has been estimated by the Panel at 3.2 and 7.1 µg/kg bw/day. Clear soft drinks contribute insignificantly with 0.001 and 0.006 µg/kg bw/day, respectively. For children, exposure is estimated at 5.7 and 13 µg/kg bw/day for the average and high level (95 th percentile) intakes, respectively. In 2

addition, according to the petitioner the vinyl esters of the fatty acids are hydrolysed in the gastrointestinal tract, the vinyl portion of the fatty acid vinyl ester instantly tautomerises to acetaldehyde and can only be detected in food in the form of acetaldehyde. The Panel concludes that the exposure to the vinyl portion of the vinyl esters of fatty acids, if present, will be very low. The maximum residual level of acetaldehyde in sucrose esters of fatty acids, as reported by the petitioner, is 48.3 mg/kg. The daily average and high level (95 th percentile) consumer exposure for Irish adults to acetaldehyde from all foodstuffs in which sucrose esters of fatty acids are permitted has been estimated by the Panel at 1.4 and 3.1 µg/kg bw/day, respectively. For children, exposure is estimated at 2.5 and 5.7 µg/kg bw/day for the average and high level (95 th percentile) consumer intakes, respectively. Clear soft drinks among adults contribute marginally with 0.0005 and 0.003 µg/kg bw, respectively. Although the International Agency for Research on Cancer (IARC) recently concluded when assessing alcohol that acetaldehyde associated with alcohol consumption is carcinogenic to humans (Group 1), the Panel noted that the evaluation by IARC was mainly based on experimental data obtained from animals after inhalation exposure and on human epidemiological data considering polymorphisms of the enzymes involved in ethanol metabolism, i.e. alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), while in the single available carcinogenicity study in which the animals were orally exposed to acetaldehyde the effects were not dose-related and no clear conclusion could be drawn from this study. In the light that acetaldehyde occurs naturally in many fruits and vegetables and other food categories, e.g. up to 132 mg/kg in orange juice and up to 10 mg/kg in bread, and that it can occur endogenously in blood plasma resulting from metabolism of ethanol and carbohydrates, the Panel considered that an additional average exposure of 1.4 µg/kg bw/day resulting from the use of sucrose esters of fatty acids as food additive would be negligible and not of safety concern. The Panel notes that the maximum residual level of p-methoxyphenol in sucrose esters of fatty acids as reported by the petitioner was <0.1 mg/kg and concludes that any exposure to this impurity resulting from the use of sucrose esters of fatty acids as food additive would be negligible and not of safety concern. The Panel therefore concludes that sucrose esters of fatty acids produced by the new manufacturing method do not present any safety concern provided the exposure is within the ADI of 40 mg/kg bw/day for sucrose esters of fatty acids and sucroglycerides. However, the Panel notes that the EC specifications for sucrose esters of fatty acids may need to be amended to include the sucrose ester of lauric acid and to permit supercritical carbon dioxide as an approved solvent to be used for their preparation. The Panel additionally concludes that an additional use of sucrose esters of fatty acids in fruit beverages only contributes to a few percent of the group ADI of 40 mg/kg bw/day for sucrose esters of fatty acids (E 473) and sucroglycerides (E 474) established by EFSA in 2004. The Panel notes however that Tier 2 intake estimates calculated for Irish consumers give a mean dietary exposure to sucrose esters of fatty acids of 29.1 mg/kg bw/day, and of 64.2 mg/kg bw/day at the 95 th percentile, the intake by high level adult consumers being above the ADI. The Panel notes that the mean dietary exposure of Irish children aged 5-12 years was 51.6 mg/kg bw/day, and 117.3 mg/kg bw/day at the 95 th percentile. For both groups (average intake and high level consumers), the estimated intakes were above the ADI. The Panel noted that in the refined estimates the main contribution to total mean dietary exposure was from fruits (36% and 46% for adults and children, respectively), due to the use of sucrose esters of fatty acids as a surface glazing agent. For a more refined estimate, exact usage data of sucrose esters of fatty acids in fruit would have to be known, or in absence of the former, information on the exact depth of the application layer, the types of fruit which are treated with the glazing agent and the 3

market share of the glazing agent in comparison to other glazing agents/waxes would have to be provided. 4

TABLE OF CONTENTS Abstract... 1 Key words... 1 Summary... 2 Table of contents... 5 Background as provided by the European Commission... 6 Terms of reference as provided by the European Commission... 6 Assessment... 7 1. Introduction... 7 2. Technical data... 7 2.1. Identity of the substance... 7 2.2. Specifications... 8 2.3. Manufacturing process... 8 2.4. Methods of analysis in foods... 9 2.4.1. Impurities in finished products... 9 2.4.2. Analysis of foods... 9 2.5. Reaction and fate in food... 9 2.6. Case of need and proposed uses... 10 2.7. Information on existing authorisations and evaluations... 11 2.7.1. Sucrose esters of fatty acids... 11 2.7.2. Acetaldehyde... 11 2.8. Exposure... 12 2.8.1. Exposure assessment for sucrose esters of fatty acids... 12 2.8.1.1. Actual levels of use of sucrose esters of fatty acids... 12 2.8.1.2. Exposure assessment... 14 2.8.2. Exposure assessment for vinyl fatty acid esters... 19 2.8.3. Exposure assessment for acetaldehyde... 19 2.8.4. Exposure assessment for p-methoxyphenol... 20 3. Biological and toxicological data... 21 3.1. Summary of the toxicological data of the EFSA opinion on sucrose esters of fatty acids (EFSA, 2004, modified on 25 January 2006)... 21 3.2. Lauric acid... 22 3.2.1. Absorption, distribution, metabolism and excretion (ADME)... 22 3.2.1.1. In vitro studies... 22 3.2.1.2. Animal studies... 22 3.2.1.3. Human studies... 22 3.2.2. Toxicological data... 22 3.2.2.1. Acute toxicity... 22 3.2.2.2. Subchronic toxicity... 23 3.2.2.3. Genotoxicity... 23 3.2.2.4. Chronic toxicity... 23 3.2.2.5 Other studies... 23 3.3. Acetaldehyde and the vinyl portion of the vinyl fatty acid... 24 3.3.1. Vinyl fatty acid esters... 24 3.3.1.1. In vitro studies... 24 3.3.2. Acetaldehyde... 24 3.4. p-methoxyphenol... 25 4. Discussion... 26 Conclusions... 28 Documentation provided to EFSA... 29 References... 29 Glossary and abbreviations... 35 5

BACKGROUND AS PROVIDED BY THE EUROPEAN COMMISSION Sucrose esters of fatty acids were evaluated by the EC Scientific Committee for Food (SCF) in 1992. At that time the SCF established a group ADI of 0-20 mg/kg bw (expressed as sucrose monostearate) for sucrose esters of fatty acids and sucroglycerides derived from palm oil, lard and tallow fatty acids. In 2004, in light of new studies which had been provided, the European Food Safety Authority (EFSA) re-examined the safety of these food additives and established a group ADI of 40 mg/kg bw/day for sucrose esters of fatty acids (E 473) and sucroglycerides (E 474). Sucrose esters of fatty acids (E 473) are authorised for use in a number of foods via Directive 95/2/EC of the European Parliament and Council on food additives other than colours and sweeteners. The specific purity criteria for sucrose esters are laid down in Commission Directive 2008/84/EC which defines sucrose esters as 'essentially the mono-, di and triesters of sucrose with fatty acids occurring in food fats and oils. They may be prepared from sucrose and methyl and ethyl esters of food fatty acids. The European Commission has now received a request to additionally authorise sucrose esters which are produced from an alternative route by reacting sucrose and vinyl esters of food fatty acids. In addition the applicant has requested an extension of use of sucrose esters to allow the additives to be used in flavourings in order to improve solubility of the flavouring in water based beverages. TERMS OF REFERENCE AS PROVIDED BY THE EUROPEAN COMMISSION In accordance with Article 29 (1) (a) of Regulation (EC) No 178/2002, the European Commission asks the European Food Safety Authority to provide a scientific opinion on the safety of sucrose esters of fatty acids prepared from vinyl esters of fatty acids. In addition, EFSA is asked to issue an opinion on the additional proposed use of sucrose esters of fatty acids in flavourings. 6

ASSESSMENT 1. Introduction The present opinion deals with the safety in use as food additive of sucrose esters of fatty acids (lauric acid, myristic acid, palmitic acid and stearic acid) produced by reacting sucrose and vinyl esters of fatty acids. The sucrose esters of all of these fatty acids, other than that of lauric acid, have previously been evaluated by the European Food Safety Authority (EFSA) (EFSA, 2004). In addition to the existing uses for sucrose esters of fatty acids, the applicant is requesting an extended use as an emulsifier to enable flavouring oils to be added to water-based beverages. The opinion therefore also considers the impact of this extended use on the total intake of sucrose esters of fatty acids. 2. Technical data 2.1. Identity of the substance Sucrose esters of fatty acids consist of a mixture of essentially mono-, di- and tri-esters of sucrose with food fatty acids. The fatty acids form esters with the hydroxyl groups in the sucrose molecule and in principle one sucrose molecule could accommodate a total of eight fatty acid molecules. Depending on the number of esterified fatty acids on a sucrose molecule, commercial products are classified in five groups as shown in Table 1. Table 1: Classification of sucrose esters of fatty acids Group Composition of esters % Specifications Sucroglycerides (E 474) Sucrose esters of fatty acids (E 473) Sucrose oligoesters type II (INS 473a) Sucrose oligoesters type I (INS 473b) Olestra (authorised in USA as a fat substitute) mono- to tri-esters 40-60% (contain residual mono-, di- and triglycerides) mono- to tri-esters 80% mono- to octa-esters 20-80%, heptaand octa-esters 20%, octa-esters 10% tetra- to octa-esters 20-80%, heptaand octa-esters 50%, octa-esters 20 Commission Directive 2008/84/EC 4 ; JECFA, 1997 Commission Directive 2008/84/EC; JECFA, 2007 JECFA, 2009 JECFA, 2009 hexa-, hepta- and octa-esters 97% FCC VI, 2008 Sucrose esters of fatty acids are non-ionic surfactants consisting of sucrose as a hydrophilic group and fatty acids as lipophilic groups. The lipophilic character increases with an increasing degree of esterification and the nature of the fatty acid in the ester group. The chemical specifications of the fatty acids in the application specify the content to be not less than 80% of sucrose monolaurate (CAS No. 25339-99-5), sucrose monomyristate (CAS No 27216-47-3), sucrose monopalmitate (CAS No. 26446-38-8), and sucrose monostearate (CAS No. 25168-73-4), with the following structural formula. 4 Commission Directive 2008/84/EC of 27 August 2008 laying down specific purity criteria on food additives other than colours and sweeteners. Official Journal of the European Communities, L 253, 20.9.2008, p. 107-108 7

CH 2 OOCR HO O OH OH O CH 2 OH O HO CH 2 OH OH R = C n H 2n+1 (n = 11, 13, 15 or 17) Figure 1: Structural formula of sucrose monoesters 2.2. Specifications Specifications for sucrose esters of fatty acids and sucroglycerides are provided in the Commission Directive 2008/84/EC laying down specific purity criteria on food additives other than colours and sweeteners. The most recent specifications for sucrose esters of fatty acids (not less than 80% sucrose esters) have been prepared by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) at its 68 th meeting in 2007 (JECFA, 2007). According to the applicant, the purity criteria as set out in Directive 2008/84/EC and the specifications set out by JECFA for sucrose esters of fatty acids are achieved when the additive is produced by the new manufacturing process using vinyl esters of fatty acids. However, according to the submitted data for several lots of the commercial products, the manufacturing process results in low levels of vinyl fatty acid residues (from <10 mg/kg to 111.23 mg/kg, limit of quantification (LOQ) between 5 and 10 mg/kg), acetaldehyde (from 17.36 mg/kg to 48.29 mg/kg, LOQ 5 µg/kg) and p-methoxyphenol (<100 µg/kg, LOQ 100 µg/kg). The petitioner proposed to include in the specifications an additional solvent, supercritical carbon dioxide, in the list of approved solvents to be used for the preparation of sucrose esters of fatty acids. The Panel notes that due to the high volatility of supercritical carbon dioxide, residues of this solvent in the final product will be extremely low. The Panel notes that the EC specifications for sucrose esters of fatty acids may need to be amended to include the sucrose ester of lauric acid and to permit supercritical carbon dioxide as an approved solvent to be used for their preparation. 2.3. Manufacturing process According to the Commission Directive 96/77/EC 5, sucrose esters of fatty acids are either prepared from sucrose and the methyl and ethyl esters of food fatty acids or by extraction from sucroglycerides. No organic solvent other than dimethylsulphoxide, dimethylformamide, ethyl acetate, propane-2-ol, 2- methyl-1-propanol, propylene glycol and methyl ethyl ketone may be used for their preparation. The new method of manufacturing sucrose esters of fatty acids is different from the existing production method. The starting fatty acid esters used are vinyl esters rather than the traditional methyl and ethyl esters of fatty acids. The initial starting material also contains a stabiliser, p-methoxyphenol. This change in manufacturing method results in trace levels of other residual chemicals, e.g. vinyl fatty acid residues, acetaldehyde (formed from the vinyl portion of the vinyl fatty acid esters) and p- methoxyphenol, not found in sucrose esters of fatty acids manufactured by the traditional methods. The reaction sequence involves the initial reaction of sucrose with the fatty acid esters followed by a series of purification steps to remove reaction solvents and the fatty acid vinyl ester starting materials. 5 Commission Directive 96/77/EC of 2 December 1996 laying down specific purity criteria on food additives other than colours and sweeteners. Official Journal of the European Communities, L 339, 30.12.1996, p. 97 8

The solvent is removed by distillation and the residue mixed with water. The mixture is sprayed into supercritical CO 2 to remove solvent residues. 2.4. Methods of analysis in foods 2.4.1. Impurities in finished products Residue levels of unreacted vinyl esters have been determined in the finished products by dissolution of the sugar esters in butanol or methanol and direct analysis by Gas Chromatography (GC) using a Flame Ionisation Detector (FID). The LOQs of this method were demonstrated to be 10, 5 and 5 mg/kg for vinyl laurate, vinyl palmitate and vinyl stearate, respectively. Residue levels of acetaldehyde have been determined by reaction with 2,4-dinitrophenylhydrazine and High Performance Liquid Chromatography (HPLC) determination of the hydrazone product with Ultraviolet (UV) detection. This method was shown to have an LOQ of 0.005 mg/kg. Residue levels of p-methoxyphenol polymerisation inhibitor have been determined by a non-specific activation test based on foaming with a claimed LOQ of 0.1 mg/kg. 2.4.2. Analysis of foods A method has been published for estimating the total content of E 473 (sucrose esters of fatty acids) in foods (Scotter et al., 2006). After selective solvent extraction of the food to recover the intact esters, they were hydrolysed to liberate sucrose, and then after acidic hydrolysis and silylation, gas chromatography/mass spectroscopy (GC/MS) was used to separately determine the glucose and fructose. The method determines the total sucrose esters content of a food sample and does not attempt to discriminate between individual sucrose esters when present as a mixture in a food sample. The method was applied to the analysis of eight different food types (including bakery wares, sugar confectionery, dairy product, margarine, meat pies and a sauce) and was shown to have a limit of quantification of around 50 mg/kg with an average recovery of 91%. The precision of the method (the relative standard deviation, RSD) averaged 11%. 2.5. Reaction and fate in food The stability, reaction and fate in food of sucrose esters of fatty acids have been evaluated and described in previous evaluations by the SCF (1992), JECFA (1997) and EFSA (EFSA, 2004, modified on 25 January 2006). When stored at room temperature as dry powder, sucrose esters of fatty acids produced according to the new manufacturing process are expected to be stable for at least 2 years. The applicant is currently conducting a bulk stability test at 25 ºC for sucrose palmitate monoester where interim results after one year indicate an approximate 2% change in monoester content. Under conditions of use in fruit flavoured beverages that have phs in the 3.8 to 5.0 range, it is expected that some hydrolysis of the sucrose esters of fatty acids may occur resulting in the formation of small amounts of free sucrose and fatty acids. Potentially, sucrose may also hydrolyse further into glucose and fructose. Surrogate stability studies have been performed and reported for sucrose monolaurate (Anderson and Polack, 1968). Buffer solutions at various ph values (2.1 to 9.3), containing approximately 10 mm sucrose monolaurate, were heated to 100 ºC for 20 hours and analysed for free lauric acid over time. 9

Maximum stability was observed between ph 4 to 5 where approximately 6.7% hydrolysis occurred after 20 hours at 100 ºC. 2.6. Case of need and proposed uses Sucrose esters of fatty acids are approved food additives (E 473) allowed to be used in a wide range of food stuffs. The applicant wishes to market sucrose monoesters of lauric acid, myristic acid, palmitic acid and stearic acid for the currently authorised applications and in addition as emulsifiers in flavour concentrates for use in fruit flavoured beverages. The new area of use is for solubilisation of essential oils without the need for processing of the oils. Without the addition of an emulsifier the flavour oil would not be soluble in water based beverages resulting in a less even dispersal of the flavouring, an increased exposure to oxygen and reduced organoleptic acceptability. The use of sucrose esters enables clear beverages with improved sensory properties to be obtained. Table 2 provides an overview of the maximum permitted levels (MPLs) of sucrose esters of fatty acids as established by the Commission Directive 95/2/EC 6 on food additives other than colours and sweeteners. Table 2: Maximum permitted levels of sucrose esters of fatty acids in foodstuffs Food group Maximum permitted level Beverages (g/l) Canned liquid coffee 1 Non-alcoholic aniseed-based drinks 5 Non-alcoholic coconut and almond drinks 5 Spirituous beverages (excluding wine and beer) 5 Dairy-based drinks 5 Foodstuffs (g/kg) Heat-treated meat products 5 (on fat) Fat emulsions for baking purposes 10 Fine bakery wares 10 Beverage whiteners 20 Edible ices 5 Sugar confectionery 5 Desserts 5 Sauces 10 Soups and broths 2 Fresh fruits, surface treatment quantum satis Powders for the preparation of hot beverages (g/l) 10 Food supplements as defined in Directive 2002/46/EC quantum satis Chewing gum 10 Cream analogues 5 Sterilised cream and sterilized cream with reduced fat content 5 Food for particular nutritional uses (PARNUTS) Infant formula and Follow on Formula for Children in good health containing hydrolysed proteins, peptides or amino acids 120 mg/l Dietary foods for special medical purposes as defined in Commission Directive 1999/21/EC 7 ; dietetic formulae for weight control intended 5 g/kg to replace total daily food intake or an individual meal 6 European Parliament and Council Directive No 95/2/EC of 20 February 1995 on food additives other than colours and sweeteners. Official Journal of the European Communities, L 61, 18. 3. 1995, p. 1. 7 Commission Directive 1999/21/EC of 25 March 1999 on dietary foods for special medical purposes. Official Journal of the European Communities, L 91, 7. 4. 1999, p. 29 10

2.7. Information on existing authorisations and evaluations 2.7.1. Sucrose esters of fatty acids Sucrose esters of fatty acids (E 473) are approved food additives regulated by the European Parliament and Council Directive 95/2/EC 8 on food additives other than colours and sweeteners. They can be used quantum satis in food supplements and for surface treatment of fresh fruit and are authorised in a number of other foods via Maximum Permitted Levels (MPLs) varying from 1 to 20 g/kg. E 473 is authorised for use in infant formulae with a MPL of 120 mg/l in products containing hydrolysed proteins, peptides or amino acids. E 473 is authorised quantum satis for use as carrier for colours and for fat soluble antioxidants. Sucrose esters of fatty acids (and sucroglycerides) were evaluated by the SCF in 1992 and a group ADI of 20 mg/kg bw/day (expressed as sucrose monostearate) was established for sucrose esters of fatty acids and sucroglycerides derived from palm oil, lard and tallow fatty acids, providing that specifications would limit the presence of tetra and higher esters to no more than 7%. The basis for establishing the numerical ADI was not specified by the SCF and no report was issued (SCF, 1992). Sucrose esters of fatty acids (and sucroglycerides) were evaluated by JECFA in 1992 and 1995. In the later evaluation JECFA allocated a temporary group ADI of 0-20 mg/kg bw/day (WHO, 1995). JECFA made the ADI temporary and requested the results of a well designed and conducted tolerance study for review in 1997 (WHO, 1995). This study was submitted and evaluated by JECFA in 1997 and a full group ADI of 0-30 mg/kg bw/day was established (WHO, 1998). In 2009, JECFA established a group ADI of 0-30 mg/kg bw/day for sucrose esters of fatty acids, sucroglycerides and sucrose oligoesters type I and type II (JECFA, 2009). The EFSA Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food (AFC) issued an opinion on sucrose esters of fatty acid (E 473) and sucroglycerides (E 474) in 2004 (modified in 2006), establishing a group ADI of 40 mg/kg bw/day for sucrose esters of fatty acid and sucroglycerides (EFSA, 2004, modified on 25 January 2006). The sucrose monoesters of lauric acid, palmitic acid and stearic acid, produced by the new manufacturing method, are according to the American Food and Drug Administration (FDA) Generally Recognized as Safe (GRAS) for the use as emulsifiers in flavour concentrates for use in fruit flavoured beverages (Federal Register, 2008). 2.7.2. Acetaldehyde The Panel has included information on the existing evaluations and authorisations of acetaldehyde, as an impurity in sucrose esters of fatty acids produced by the new manufacturing method. Acetaldehyde originates from the vinyl alcohol portion of the vinyl esters of fatty acids, that is formed via hydrolysis upon reaction with sucrose. Acetaldehyde [FL-no 05.001] may be used as a flavouring substance according to Commission Decision 1999/217/EC adopting a register of flavouring substances used in or on foodstuffs 9, last amended by Commission Decision 2009/163/EC 10,. 8 European Parliament and Council Directive No 95/2/EC of 20 February 1995 on food additives other than colours and sweeteners. Official Journal of the European Communities, OJ L 61, 18.3.1995, p. 1 9 Commission Decision 1999/217/EC of 23 February 1999 adopting a register of flavouring substances used in or on foodstuffs drawn up in application of Regulation (EC) No 2232/96 of the European Parliament and of the Council of 28 October 1996. Official Journal of the European Communities, L 84, 27.3.1999 11

The use of acetaldehyde as a flavouring substance was evaluated by JECFA in 1997. The Committee estimated that the intakes of acetaldehyde at levels of 9.7 and 11 mg per person per day in the USA and Europe, respectively would not present safety concern (WHO, 1998). According to Commission Directive 2002/72/EC 11 acetaldehyde may be used as monomer or starting substance in the manufacture of plastic materials and articles intended to come into contact with foodstuffs. Its specific migration limit (SML) in food or in food simulant expressed as total of moiety or substance(s) indicated (SML(T)) is 6 mg/kg (Annex II of Directive 2002/72/EC). SML(T) in this specific case means that the restriction shall not be exceeded by the sum of the migration of the following substances mentioned as Ref. Nos: 10060 [acetaldehyde] and 23920 [Propionic acid, vinyl ester]. The SCF evaluated the use of acetaldehyde in the manufacture of plastic materials and articles intended to come into contact with foodstuffs and derived for acetaldehyde and vinyl propionate a group tolerable daily intake (TDI) of 0.1 mg/kg bw (SCF, 1998). Acetaldehyde as a chemical was last evaluated by the IARC in 1999 and classified in Group 2B (IARC 1999). In 2009, IARC updated their assessments of several personal habits and household exposures that cause cancer, including consumption of alcoholic beverages and the resulting exposure to acetaldehyde. Based on studies showing a higher risk of alcohol related cancer of oesophagus and head and neck in East-Asian populations that accumulate acetaldehyde formed endogenously and after consumption of alcohol, due to a deficit (genetic polymorphism) in aldehyde dehydrogenase, it was concluded that acetaldehyde associated with alcohol consumption is carcinogenic to humans (Group 1) (IARC, 2009; Secretan et al., 2009). 2.8. Exposure The exposure assessments presented in this opinion cover two aspects: (1) exposure to the sucrose esters of fatty acids themselves, from all permitted uses, including the additional use requested by the petitioner, for solubilisation of essential oils without the need for processing of the oils; (2) exposure to the potential by-products of the new manufacturing process, namely low levels of vinyl fatty acid residues, acetaldehyde (formed from the vinyl portion of the vinyl fatty acid esters) and p- methoxyphenol (stabiliser in the vinyl fatty acid starting material). 2.8.1. Exposure assessment for sucrose esters of fatty acids 2.8.1.1. Actual levels of use of sucrose esters of fatty acids More information on current use levels was made available to the Panel for several food categories in finished products. Beverages Sucrose esters of fatty acids are permitted for use in non-alcoholic aniseed-based drinks, non-alcoholic coconut and almond drinks, spirituous beverages (excluding wine and beer), milk based drinks and canned liquid coffee. In France, a survey carried out by AFSSA (Bemrah et al., 2008) on the occurrence of sucrose esters of fatty acids in a number of food categories included 13 samples of alcoholic beverages, none of which were found to contain the additive. For the new proposed application, a maximum usage level of 0.03 g/l in clear/colourless carbonated beverages (non fruit based beverages based on citrus oils, e.g. lemonades) has been indicated by the applicant. This is based on a maximum dosage in the flavouring concentrate of 1.5%. The flavouring is intended to be 10 Commission Decision 2009/163/EC of 26 February 2009 amending Decision 1999/217/EC as regards the register of flavouring substances used in or on foodstuffs (notified under document number C(2009) 1222). Official Journal of the European Communities, L 55, 27.2.2009, p. 41 11 Commission Directive 2002/72/EC of 6 August 2002 relating to plastic materials and articles intended to come into contact with foodstuffs. Journal of the European Communities, L 220, 15.8.2002, p. 18 12

incorporated into the beverage at 0.2%, giving a maximum level of sucrose esters of fatty acids in the beverage of 0.03 g/l. No actual usage or concentration data was available for any of the remaining beverage categories. Foodstuffs For fine bakery wares, the Confederation of the Food and Drink Industries (CIAA) in the EU (2009) reported a range of typical use levels of sucrose esters of fatty acids ranging from 0.4-2.3 g/kg and extreme usage values of 5 g/kg. In France, a survey carried out by the French Food Safety Agency (AFSSA) (Bemrah et al., 2008) on the occurrence of E473 in a number of food categories included 23 samples of fine bakery wares, none of which were found to contain the additive. For sugar confectionery, the CIAA (2009) reported typical use levels of sucrose esters of fatty acids ranging from 0-5 g/kg. For foods intended for special medical purposes, the CIAA (2009) reported typical use levels of sucrose esters of fatty acids ranging from 0.03-5 g/kg. For the categories fat emulsions, beverage whiteners, sauces and edible ices the CIAA (2009) report indicated no use of sucrose esters of fatty acids in these products. In France, a survey carried out by AFSSA (Bemrah et al., 2008) on the occurrence of E473 in a number of food categories included milk based desserts (27), flavoured desserts (27), powdered hot beverages (16), and soups/broths (23), none of which were found to contain the additive. Overall, there is little information on actual usage levels of sucrose esters of fatty acids in foods or beverages. However, studies conducted in France (Bemrah et al., 2008) and the UK (Scotter et al., 2006) indicate that application of the additive is very sparse. Also, in Ireland, a database on ingredients (National Food Ingredient Database - NFID) of more than 1800 foodstuffs only identified 4 foodstuffs containing the additive. In order to refine the exposure assessment for children and adults to sucrose esters of fatty acids, quantum satis applications have to be quantified. No usage data or concentration data for the relevant two food groups (surface treatment of fresh fruits, food supplements) were however, available, and levels were estimated according to a number of assumptions. For the surface treatment of fresh fruits, a level of 18 g/kg has been applied, based on the following assumptions: The surface area of an apple amounts to approximately 113 cm 2 (4πr 2, r=3 cm). Assuming an application of 0.3 mm glazing agent onto the surface, the volume (4/3πr 3 ) of the glazing agent can be calculated as follows: Volume of apple including glazing agent volume of apple excluding glazing agent (4/3 3.14159 3.03 3 ) - (4/3 3.14159 3 3 ) = 116.5-113.1 = 3.4 cm 3 According to the applicant, the density of the powder is approximately 600 kg/m 3 (0.6 g/cm 3 ) and hence the applied amount of sucrose ester of fatty acids amounts to 2 g (per apple) following (m=ρ v; mass=density volume). Based on the weight of a medium sized apple of 112 g (EC, 2004), a total concentration of 18 g/kg can be calculated. The exposure estimates for fruit are very conservative, as it is assumed that all fruits (reported as being consumed including skin) are treated with sucrose esters of fatty acids as a glazing agent and that the calculated concentration based on apples is applicable to all fruits. The application layer of 0.3 mm is also an assumption. For a more refined estimate, exact usage data of sucrose esters of fatty acids in 13

fruit would have to be known, or in absence of the former, information on the exact thickness of the application layer, the types of fruit which are treated with the glazing agent and the market share of the glazing agent in comparison to other glazing agents/waxes would have to be provided. Table 3: Maximum reported use levels of sucrose esters of fatty acids in beverages and foodstuffs used for the refined exposure assessment Beverages Maximum reported use level (g/l) Non-alcoholic aniseed-based drinks 5 Non-alcoholic coconut and almond drinks 5 Spirituous beverages (excluding wine and beer) 5 Dairy-based drinks 5 Canned liquid coffee 1 Dairy-based drinks 5 Non alcoholic clear/colourless beverages 0.03* Foodstuffs Maximum reported use level (g/kg) Heat-treated meat products 5 (on fat) Fat emulsions for baking purposes 10 Fine bakery wares 10 Beverage whiteners 20 Edible ices 5 Sugar confectionery 5 Desserts 5 Sauces 10 Soups and broths 2 Fresh fruits, surface treatment** 18 Powders for the preparation of hot beverages 10 g/l Food supplements as defined in Directive 2002/46/EC ** 10 Chewing gum 10 Cream analogues 5 Sterilised cream and sterilized cream with reduced fat content 5 * According to applicant s information ** Estimated usage in quantum satis application For food supplements, which are marketed either as liquids or in powder form, a concentration of 10 g/kg was assumed based on the similarity of the product to the matrix powders for the preparation of hot beverages. To take into account the potential additional exposure due to the new use proposed by the applicant, a maximum level of sucrose ester of 0.03 g/l in accordance with the applicant s information has been allocated to clear/colourless carbonated beverages. 2.8.1.2. Exposure assessment The Panel agreed to follow the principles of the stepwise approach, which were used in the report of the Scientific Co-operation (SCOOP) Task 4.2, to estimate additives intakes (EC, 1997). In the tiered 14

approach, Tier 1 is based on theoretical food consumption data and maximum permitted use levels (MPLs) for additives as permitted by relevant Community legislation. The Second and Third tiers refer to assessment at the level of individual Member States, combining national data on food consumption with the maximum permitted usage levels for the additive (Tier 2) and with its actual usage patterns (Tier 3). Crude estimates (Budget method) The dietary exposure to sucrose esters of fatty acids from the MPLs was estimated using the Budget method (Tier 1), which is based on the fact that there is a physiological upper limit to the amount of food and drink (for beverages 100 ml/kg bw and for solids 25 g/kg bw), and thus of food additives, that can be consumed each day. A further assumption is that only a certain proportion of the diet is likely to contain food additives (25%). Full details on the budget method are described in the report of the SCOOP Task 4.2 (EC, 1997). For adults, the maximum permitted use level of sucrose esters of fatty acids considered for beverages was 5 g/l. The maximum permitted level considered for solid foods was 20 g/kg (Table 2). The default proportion (25%) of beverages and solid food that could contain the additive was considered adequate. In effect, even though sucrose esters of fatty acids may be used in a variety of solid foods that could represent more than 25% of processed foods, it is unlikely that a person would systematically choose all processed solid foods with the same emulsifier added. In the case of beverages, uses are reported for a limited number of beverages; however, some of these may constitute a significant proportion of liquid intake (i.e., dairy based drinks), with consumer loyalty to a single brand often being high for this category of product. The 25% proportion was therefore considered adequate also for beverages (EC, 1997). This assumes that a typical adult, weighing 60 kg, consumes daily 1.5 litres (it is assumed that only 25% of the daily consumption of 6 litres contain sucrose esters of fatty acids) of beverages and 375 g of solid foods, containing sucrose esters of fatty acids. For children, the level of sucrose esters of fatty acids considered in beverages was 5 g/l (after exclusion of alcoholic drinks), and in solid food 10 g/kg as children are unlikely to consume beverage whiteners (the food group with the highest permitted level). The default proportion of 25% in solid foods was considered adequate, however for beverages it was recognised to be inadequate, as the corresponding consumption rate of 375 ml/day could easily be exceeded by young children. This conclusion was derived from UK data on consumption of soft drinks by children aged less than 5 years, where the 97.5 th percentile of consumption was between 70 and 80 ml/kg bw/day and a proportion factor of 100 % for beverages was recommended for children in the SCOOP Task 4.2 (EC, 1997). This assumes that a typical 3-year old child, weighing 15 kg, consumes daily 1.5 litres of beverages and 94 g of solid foods. The theoretical maximum daily exposure for adults and children is summarised in Table 4. 15

Table 4: Theoretical maximum daily exposure for adults and children Daily upper physiological limit of intake per kg/bw Beverage ml/day Food g/day Amount of food/beverage assumed to contain additive (%) Beverage ml (%) Food g (%) Maximum permitted level of sucrose esters of fatty acids Beverage g/l Food g/kg Theoretical maximum intake of sucrose esters of fatty acids Beverage g/day Food g/day Beverage mg/kg bw/day Food mg/kg bw/day Adult (60 kg) 6000 1500 1500 (25) 375 (25) 5 20 7.5 7.5 125 125 250 Child (15 kg) 1500 375 1500 (100) 94 (25) 5 10 7.5 0.94 500 63 563 Total mg/kg bw/day 16

Combined exposure to sucrose esters of fatty acids from beverages and food (mg/kg bw/day): Adult: ((6000 0.25)/1000 5)/60 + ((1500 0.25)/1000 20)/60 = 250 mg/kg bw/day Child: (1500 1)/1000 5)/15 + ((375 0.25)/1000 10))/15 = 563 mg/kg bw/day It was noted that sucrose esters of fatty acids may be used quantum satis in food supplements and for the surface treatment of fresh fruits. Refined estimates Refined exposure estimates have been performed for Tier 2 using MPLs presented in Table 2 and maximum practical use levels presented in Table 3 to deal with the specific cases of quantum satis authorisation for food supplements and surface treated fruits for children and adult populations. As available quantitative information on usage levels is in line with existing MPLs, no separate Tier 3 calculations were performed as baseline data are identical to those used in Tier 2 calculations. As sucrose esters of fatty acids are permitted in a wide range of food groups, including very specific foodgroups, exposure estimates have been derived using detailed individual food consumption data available for Irish children 5-12 years old derived from the National Children s Food Survey (NCFS) (IUNA, 2005) and adults 18-65 years old (IUNA, 2001) for Tier 2/Tier 3. This approach was chosen, as no other suitable food consumption data for children were available to perform detailed intake assessment of sucrose esters of fatty acids. For adults, the EFSA Concise European Food Consumption Database, which gives access to aggregate food categories consumed in 15 European countries, was considered to provide insufficient details for the refined assessment of sucrose esters of fatty acids. Table 5 summarises the anticipated exposure of children and adults to sucrose esters of fatty acids. Total exposure to sucrose esters of fatty acids was calculated using Creme 2.0 probabilistic software (2005-2009 Creme Software Ltd) and a distributional exposure was derived for both the adult and children population in Ireland. In the case of sucrose esters of fatty acids, when considering MPLs of use (Tier 2), the mean dietary exposure of Irish children (aged 5-12 years and weighing on average 33 kg) from all foods in which sucrose esters of fatty acids are permitted was 51.6 mg/kg bw/day, and 117.3 mg/kg bw/day at the 95 th percentile. The main contributors to the total anticipated exposure to sucrose esters of fatty acids were fruit (46%), fine bakery wares (16.2%), and sauces (6.1%). Estimates calculated for the Irish adult population give a mean dietary exposure to sucrose esters of fatty acids of 29.1 mg/kg bw/day, and of 64.2 mg/kg bw/day at the 95 th percentile. The main contributors to the total anticipated exposure to sucrose esters of fatty acids were fruit (35.8%), fine bakery wares (15.0 %), and hot drinks prepared from powders (11.1%). As the above assumptions incorporate a great degree of uncertainty, results should be interpreted with the necessary caution. Refined intake estimates for the new application of sucrose esters of fatty acids to be used in flavourings in water based beverages have been estimated by the Panel based on Irish adult and children population and usage data provided by the applicant. For adults an average intake of 0.01 mg/kg bw/day and a 95 th percentile of 0.06 mg/kg bw/day, contributing 0.03% to total mean intake of sucrose esters of fatty acids, were calculated. For children, a mean of 0.04 mg/kg bw/day and high intake (95 th percentile) of 0.18 mg/kg bw/day contributing 0.1% to the total mean exposure of sucrose esters of fatty acids, was calculated. 17

Table 5: Summary of anticipated exposure to sucrose esters of fatty acids using tiered approach (EC, 2001) in children and adult populations Tier 1. Budget method Adult population Children population 250 mg/kg bw/day 562 mg/kg bw/day Food Group Tier 2. Maximum permitted level/ Tier 3. Maximum reported use levels (Total) Irish adult population (18-65) (Total) Irish children population (5-12) Mean P95 Contribution Mean P95 Contribution mg/kg bw/day mg/kg bw/day % mg/kg bw/day mg/kg bw/day % Beverages (10%) (1) 0.92 5.37 3.2 2.55 11.07 4.9 Beverages clear (2) 0.01 0.06 0.03 0.04 0.18 0.1 Milk Based Beverages 0.06 0.00 0.2 0.62 5.49 1.2 Spirituous Beverages 0.46 2.66 1.6 0.0002 0.00 0.0004 Cream sterilised and UHT 0.003 0.00 0.01 0.001 0.00 0.0019 Desserts 1.48 6.03 5.1 1.97 9.20 3.8 Edible ices 0.50 2.40 1.7 2.41 8.76 4.7 Fat Emulsions 2.58 6.80 8.9 2.62 6.60 5.1 Fine Bakery Ware (5) 4.35 12.87 15.0 8.38 21.01 16.2 Fruit (3) 10.41 40.38 35.8 23.78 80.63 46.1 Fat from Processed meat 0.42 1.18 1.4 0.68 2.06 1.3 Sauces (4) 2.33 7.97 8.0 3.17 10.05 6.1 Soups (4) 0.69 3.06 2.4 0.73 4.45 1.4 Sugar Confectionery 0.10 0.65 0.3 2.02 7.54 3.9 Coffee Whitener 0.001 0.00 0.004 0.00 0.00 0.0 Hot Drink Powders (5) 3.23 3.77 11.1 1.52 9.33 2.9 Supplements (6) 1.50 4.00 5.2 1.00 2.00 1.9 Chewing Gum 0.01 0.00 0.0231 0.09 0.58 0.2 Total 29.1 64.2 100 51.6 117.3 100 (1) Sucrose esters of fatty acids are permitted only in non-alcoholic soft drinks based on aniseed, coconut or almond. No such products were found in the Irish Food Consumption Databases. Therefore and due to the specificity of these food categories, exposure was estimated assuming that 10% of non alcoholic beverages contain sucrose esters of fatty acids (10% presence probability). (2) Clear beverages (e.g. lemonades, excluding all cola and fruit based beverages) for which a new application of sucrose esters of fatty acids has been applied for (0.03 g/l) (3) Fruit which was reported as being consumed or used in a recipe as whole (e.g. unpeeled) (4) Excluding homemade products (5) As consumed (i.e. made up) (6) Sucrose esters of fatty acids are permitted in supplements at quantum satis. An MPL of 10 g/kg has been assumed based on the comparability of the substrate with powders for the preparation of hot drinks. As consumption of food supplements is reported in units rather than in mass, exposure could not be calculated as part of the overall distributional exposure. The average unit weight of a typical food supplement is 100mg, therefore an additional exposure of 1 g per unit can be assumed. Mean and 95 th percentile daily unit intake for adults was 1.5 and 4 units respectively and for children 1 and 2 units respectively. These were added to the distributional exposure figures. Sucrose esters of fatty acids are authorised for use in infant formulae and follow on formulae for infants in good health with a maximum permitted level of 120 mg/l in products containing hydrolysed 18